CA2210951A1 - Modified thermostable dna polymerase - Google Patents

Modified thermostable dna polymerase

Info

Publication number
CA2210951A1
CA2210951A1 CA002210951A CA2210951A CA2210951A1 CA 2210951 A1 CA2210951 A1 CA 2210951A1 CA 002210951 A CA002210951 A CA 002210951A CA 2210951 A CA2210951 A CA 2210951A CA 2210951 A1 CA2210951 A1 CA 2210951A1
Authority
CA
Canada
Prior art keywords
residue
sequence
dna
polymerases
thermostable dna
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CA002210951A
Other languages
French (fr)
Other versions
CA2210951C (en
Inventor
David Harrow Gelfand
Lisa Vivian Kalman
Fred Lawrence Reichert
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
F Hoffmann La Roche AG
Original Assignee
F Hoffmann La Roche AG
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by F Hoffmann La Roche AG filed Critical F Hoffmann La Roche AG
Publication of CA2210951A1 publication Critical patent/CA2210951A1/en
Application granted granted Critical
Publication of CA2210951C publication Critical patent/CA2210951C/en
Anticipated expiration legal-status Critical
Expired - Fee Related legal-status Critical Current

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6844—Nucleic acid amplification reactions
    • C12Q1/6848—Nucleic acid amplification reactions characterised by the means for preventing contamination or increasing the specificity or sensitivity of an amplification reaction
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1241—Nucleotidyltransferases (2.7.7)
    • C12N9/1252—DNA-directed DNA polymerase (2.7.7.7), i.e. DNA replicase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6869—Methods for sequencing
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/52—Genes encoding for enzymes or proenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Molecular Biology (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Saccharide Compounds (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)

Abstract

The invention provides thermostable DNA polymerase enzymes that comprises the amino acid sequence SerGlnIleXaaLeuArgXaa (SEQ ID NO: 1), wherein "Xaa" at position 4 of this sequence is any amino acid residue but not a glutamic acid residue (Glu), preferably a glycine residue and "Xaa" at position 7 of this sequence is a valine residue (Val) or an isoleucine residue (Ile). The thermostable DNA polymerases of the invention have enhanced efficiency for incorporating unconventional nucleotides, such as ribonucleotides, into DNA products and are advantageous in many in vitro synthesis applications. Such enzymes are particularly useful for use in nucleic acid sequencing protocols and provide novel means for DNA sequence analysis with cost and efficiency advantages. Also claimed are nucleic acids encoding said polymerases, vectors and hoste cells comprising such a nucleic acid, as well as compositions for use in a DNA sequencing reaction, kits and methods for sequencing including such polymerases.
CA002210951A 1996-08-06 1997-08-01 Modified thermostable dna polymerase Expired - Fee Related CA2210951C (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US2337696P 1996-08-06 1996-08-06
US60/023,376 1996-08-06

Publications (2)

Publication Number Publication Date
CA2210951A1 true CA2210951A1 (en) 1998-02-06
CA2210951C CA2210951C (en) 2009-02-10

Family

ID=21814728

Family Applications (1)

Application Number Title Priority Date Filing Date
CA002210951A Expired - Fee Related CA2210951C (en) 1996-08-06 1997-08-01 Modified thermostable dna polymerase

Country Status (21)

Country Link
US (1) US5939292A (en)
EP (1) EP0823479B1 (en)
JP (1) JP3170229B2 (en)
KR (1) KR100510619B1 (en)
CN (1) CN1154733C (en)
AT (1) ATE278020T1 (en)
AU (1) AU714929B2 (en)
BR (1) BR9704260B1 (en)
CA (1) CA2210951C (en)
CZ (1) CZ293215B6 (en)
DE (1) DE69730926T2 (en)
DK (1) DK0823479T3 (en)
ES (1) ES2227640T3 (en)
HU (1) HUP9701341A3 (en)
IL (1) IL121441A (en)
MX (1) MX9705961A (en)
NO (1) NO322135B1 (en)
PL (1) PL321478A1 (en)
RU (1) RU2235773C2 (en)
TW (1) TW528802B (en)
UA (1) UA47423C2 (en)

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US6458945B1 (en) 1998-10-01 2002-10-01 Variagenics, Inc. Method for analyzing polynucleotides
US6777188B2 (en) 1998-10-01 2004-08-17 Variagenics, Inc. Genotyping by mass spectrometric analysis of allelic fragments
US6855500B2 (en) 1998-10-01 2005-02-15 Sequenom, Inc. Fluorescence-based genotyping
US6994998B1 (en) 1998-10-01 2006-02-07 Sequenom, Inc. Base-modified nucleotides and their use for polymorphism detection
US6500650B1 (en) 1998-10-01 2002-12-31 Variagenics, Inc. Method for identifying polymorphisms
US6610492B1 (en) 1998-10-01 2003-08-26 Variagenics, Inc. Base-modified nucleotides and cleavage of polynucleotides incorporating them
US6440705B1 (en) 1998-10-01 2002-08-27 Vincent P. Stanton, Jr. Method for analyzing polynucleotides
AU5008900A (en) * 1999-05-12 2000-11-21 Invitrogen Corporation Compositions and methods for enhanced sensitivity and specificity of nucleic acid synthesis
GB9921318D0 (en) * 1999-09-09 1999-11-10 Kristensen Tom Chimeric molecules
US6329178B1 (en) 2000-01-14 2001-12-11 University Of Washington DNA polymerase mutant having one or more mutations in the active site
US7179590B2 (en) 2000-04-18 2007-02-20 Roche Molecular Systems, Inc High temperature reverse transcription using mutant DNA polymerases
US6214557B1 (en) * 2000-06-06 2001-04-10 Washington University Cold sensitive mutant DNA polymerases
US20020102577A1 (en) * 2000-07-31 2002-08-01 Raillard Sun Ai Nucleotide incorporating enzymes
US6811979B2 (en) * 2000-10-11 2004-11-02 Applera Corporation Fluorescent nucleobase conjugates having anionic linkers
IL155450A0 (en) * 2000-10-20 2003-11-23 Expression Diagnostics Inc Leukocyte expression profiling
WO2002073504A1 (en) * 2001-03-14 2002-09-19 Gene Logic, Inc. A system and method for retrieving and using gene expression data from multiple sources
US7026121B1 (en) 2001-06-08 2006-04-11 Expression Diagnostics, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US6905827B2 (en) 2001-06-08 2005-06-14 Expression Diagnostics, Inc. Methods and compositions for diagnosing or monitoring auto immune and chronic inflammatory diseases
US7235358B2 (en) 2001-06-08 2007-06-26 Expression Diagnostics, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US6887690B2 (en) * 2001-06-22 2005-05-03 Pe Corporation Dye-labeled ribonucleotide triphosphates
CA2460546A1 (en) * 2001-09-14 2003-03-27 Invitrogen Corporation Dna polymerases and mutants thereof
US20050042629A1 (en) 2002-09-13 2005-02-24 Applera Corporation Thermus scotoductus nucleic acid polymerases
WO2003048308A2 (en) 2001-11-30 2003-06-12 Applera Corporation Thermus brockianus nucleic acid polymerases
US6723546B2 (en) * 2002-03-26 2004-04-20 New England Biolabs, Inc. Method for cloning and expression of BsaI restriction endonuclease and BsaI methylase in E. coli
US7148049B2 (en) 2002-04-02 2006-12-12 Roche Molecular Systems, Inc. Thermostable or thermoactive DNA polymerase molecules with attenuated 3′-5′ exonuclease activity
US7820030B2 (en) 2003-04-16 2010-10-26 Handylab, Inc. System and method for electrochemical detection of biological compounds
US7892745B2 (en) 2003-04-24 2011-02-22 Xdx, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US7572581B2 (en) * 2003-06-30 2009-08-11 Roche Molecular Systems, Inc. 2′-terminator nucleotide-related methods and systems
US7947817B2 (en) * 2003-06-30 2011-05-24 Roche Molecular Systems, Inc. Synthesis and compositions of 2'-terminator nucleotides
EP1493824A1 (en) * 2003-07-02 2005-01-05 Consortium National de Recherche en Genomique (CNRG) Method for detection of mutations in DNA
JP4127204B2 (en) * 2003-12-17 2008-07-30 セイコーエプソン株式会社 Manufacturing method of liquid crystal display device
US20080311627A1 (en) * 2004-06-17 2008-12-18 Epigenomics Ag Compositions and Methods for Preventing Carry-Over Contamination in Nucleic Acid Amplification Reactions
US7745125B2 (en) 2004-06-28 2010-06-29 Roche Molecular Systems, Inc. 2′-terminator related pyrophosphorolysis activated polymerization
US7645575B2 (en) 2004-09-08 2010-01-12 Xdx, Inc. Genes useful for diagnosing and monitoring inflammation related disorders
US7890268B2 (en) * 2004-12-28 2011-02-15 Roche Molecular Systems, Inc. De-novo sequencing of nucleic acids
US20060292578A1 (en) * 2005-06-28 2006-12-28 Weidong Zheng DNA polymerase blends and mutant DNA polymerases
WO2007002890A2 (en) * 2005-06-28 2007-01-04 Agencourt Personal Genomics Corporation Methods of producing and sequencing modified polynucleotides
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US20070154894A1 (en) * 2005-12-30 2007-07-05 Affymetrix, Inc. Labeling and non-enzymatic fragmentation of cDNA using a ribonucleoside triphosphate analog
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WO2008140484A2 (en) 2006-11-09 2008-11-20 Xdx, Inc. Methods for diagnosing and monitoring the status of systemic lupus erythematosus
US20080242560A1 (en) * 2006-11-21 2008-10-02 Gunderson Kevin L Methods for generating amplified nucleic acid arrays
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Also Published As

Publication number Publication date
IL121441A (en) 2000-07-16
BR9704260B1 (en) 2010-09-08
PL321478A1 (en) 1998-02-16
CZ293215B6 (en) 2004-03-17
EP0823479A3 (en) 2000-03-29
HU9701341D0 (en) 1997-09-29
NO973595L (en) 1998-02-09
CN1154733C (en) 2004-06-23
NO322135B1 (en) 2006-08-21
UA47423C2 (en) 2002-07-15
TW528802B (en) 2003-04-21
JP3170229B2 (en) 2001-05-28
DE69730926T2 (en) 2006-02-23
EP0823479A2 (en) 1998-02-11
CZ229997A3 (en) 1998-06-17
DE69730926D1 (en) 2004-11-04
KR100510619B1 (en) 2005-12-21
HUP9701341A3 (en) 2000-12-28
MX9705961A (en) 1998-02-28
ES2227640T3 (en) 2005-04-01
RU2235773C2 (en) 2004-09-10
JPH1066588A (en) 1998-03-10
AU714929B2 (en) 2000-01-13
IL121441A0 (en) 1998-01-04
DK0823479T3 (en) 2005-01-31
US5939292A (en) 1999-08-17
NO973595D0 (en) 1997-08-05
BR9704260A (en) 1998-09-15
AU3319797A (en) 1998-02-12
EP0823479B1 (en) 2004-09-29
CA2210951C (en) 2009-02-10
KR19980018395A (en) 1998-06-05
HUP9701341A2 (en) 1999-06-28
CN1177640A (en) 1998-04-01
ATE278020T1 (en) 2004-10-15

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Effective date: 20170801